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Characterization of the Vibrio vulnificus 1-Cys Peroxiredoxin Prx3 and Regulation of Its Expression by the Fe-S Cluster Regulator IscR in Response to Oxidative Stress and Iron Starvation

Cited 26 time in Web of Science Cited 28 time in Scopus
Authors

Lim, Jong Gyu; Bang, Ye-Ji; Choi, Sang Ho

Issue Date
2014-12
Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Citation
JOURNAL OF BIOLOGICAL CHEMISTRY, Vol.289 No.52, pp.36263-36274
Keywords
ESCHERICHIA-COLIHYDROGEN-PEROXIDEDISTINCT CHARACTERISTICS2-CYS PEROXIREDOXINSCATALYTIC MECHANISMANTIOXIDANT PROTEINASSEMBLY PROTEINSREACTIVE OXYGENFAMILYGENE
Abstract
Peroxiredoxins (Prxs) are ubiquitous antioxidant enzymes that reduce toxic peroxides. A new Vibrio vulnificus Prx, named Prx3, was identified and characterized in this study. Biochemical and mutational analyses revealed that Prx3 reduces H2O2, utilizing glutaredoxin 3 (Grx3) and glutathione (GSH) as reductants, and requires only N-terminal peroxidatic cysteine for its catalysis. These results, combined with the monomeric size of Prx3 observed under non-reducing conditions, suggested that Prx3 is a Grx3/GSH-dependent 1-Cys Prx and oxidized without forming intermolecular disulfide bonds. The prx3 mutation impaired growth in the medium containing peroxides and reduced virulence in mice, indicating that Prx3 is essential for survival under oxidative stress and pathogenesis of V. vulnificus. The Fe-S cluster regulator IscR activates prx3 by direct binding to a specific binding sequence centered at -44 from the transcription start site. The binding sequence was homologous to the Type 2 IscR-binding sequence, most likely recognized by the Fe-S clusterless apo-IscR in Escherichia coli. The iscR(3CA) mutant, chromosomally encoding the apo-locked IscR, exhibited 3-fold higher levels of activation of prx3 than the wild type and accumulated more IscR(3CA) protein in cells. The IscR-dependent activation of prx3 by aerobic growth and iron starvation was also associated with the increase in cellular levels of IscR protein. Taken together, the results suggested that IscR senses iron starvation as well as reactive oxygen species and shifts to the apo-form, which leads to the increase of cellular IscR and in turn prx3 expression, contributing to the survival and virulence of V. vulnificus during pathogenesis.
ISSN
0021-9258
URI
https://hdl.handle.net/10371/191812
DOI
https://doi.org/10.1074/jbc.M114.611020
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  • College of Medicine
  • Department of Medicine
Research Area Host-microbe Interaction, Bacterial Pathogenesis, Microbiome, Mucosal Immunity, Nutritional Immunology

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